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AssayShort Reference or Description of MethodFull Reference
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Acid Detergent FiberAnkom Technologies Method 5https://www.ankom.com/sites/default/files/document-files/Method_5_ADF_A200.pdf
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AmmoniaFermentation products are extracted from the sample in a 1:10 ratio of sample and deionized water. The extract is centrifuged and the supernatant is combined with calcium hydroxide and copper sulfate, centrifuged again, and the supernatant is analyzed by Skalar San++ Segmented Flow SA 5000 Analyzer. 

Skalar Method: Ammonia: Automated procedure  based on the modified Berthelot Reaction:  after dialysis, the ammonia is buffered and chlorinated to monochloramine which reacts with salicylate to form 5-aminosalicylate. After Oxidation and oxidative coupling  a green colored complex is formed. The absorption of the formed complex is measured at 660 nm.		Skalar Method Resources: 
ISO 3693:1987, Water for analytical laboratory use. Specification and test methods. 
ASTM, D1193, Standard Specification for Reagent Water. 
Krom,M.,”Spectrophotometric determination of ammonia; a study of modified Berthelot reaction using salicylate and dichloroisocyanurate”, The Analyst, April 1980, Vol. 105, page 305-316.
Searle,P.L., “The Berthelot or indophenol reaction and its use in the analysis chemistry of nitrogen”, The Analyst,Vol. 109, May 1984, page 549-565
EN ISO 11732, Determination of ammonium by flow analysis and spectrometric determination.
ISO 3696:1987, Water for analytical laboratory use. Specification and test methods. 
ASTM, D1193, Standard Specification for Reagent Water.  
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Ash(AOAC 942.05)
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ChlorideSample is extracted with 50 mL of distilled water and analyzed by titration with silver chloride using a Nelson-Jameson M926S chloride analyzer. Nelson-Jameson Inc.,3200 S. Central Ave., Marshfield WI 54449
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ClostridiumClostridium Perfringens by Membrane Filtration using CP Chromoselect Agar.
A 10g blended subsample is diluted using butterfields buffer, plated on CP Chromoselect Agar (12398, MilliporeSigma) , and incubated at 44 C for 24 hours in an anaerobic chamber. Visible colonies are counted , multiplied by 10 to account for the dilution, and reported.
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Crude Protein(AOAC 990.03)Our crude protein is determined by Dumas combustion and Nitrogen determination. (N * 6.25)
We have a Rapid N analyzer by Elementar and a Leco True Mac.
AOAC Official Method 990.03 (2005) Official Methods of
Analysis of AOAC INTERNATIONAL, 18th Ed., AOAC
INTERNATIONAL, Gaithersburg, MD
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Dry MatterNFTA AddendumShreve, B., & Thiex, N. 2006. NFTA Method 2 . 1 . 4 - Dry Matter by Oven Drying for 3 hr at 105 C NFTA Method 2 . 1 . 4 - Dry Matter by Oven Drying for 3 hr at 105 C, (January), 4–7.
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Fatty Acid ProfileFatty acids in feeds go through a methylation procedure to produce Fatty Acid Methyl Esters of the desired Fatty Acids. Following methylation they are extracted with ethyl ether and analyzed by GC FID. Modified from Jenkins 1-Step Methylation procedure for Fatty Acids by GC FID
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In Situ 7 h Starch Digestion3 g dried and 6mm ground sample are weighed into R150 concentrate bags (Ankom Technologies) in triplicate. Bags are placed into weighted laundry bags and inserted into 3 cannulated lactating cows at the University of Wisconsin-Madison fed a conventional lactating diet for 7 hours. Digested bags are rinsed 3 times in cold water, dried overnight, and weighed back. Residues are compiled, ground to 1 mm and run through the starch procedure (Hall, 2008).
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In Situ 16 h CP Digestion 3 g dried sample weighed into R150 concentrate bags (Ankom Technologies) in triplicate. Bags are placed into laundry bags and inserted into 3 cannulated lactating cows at the University of Wisconsin-Madison fed a conventional high fiber diet for 16 hours. Digested bags are rinsed 3 times in cold water, dried overnight, and weighed back. Residues are compiled, and run through the CP procedure (AOAC 990.03)
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In Vitro Starch DigestionFeed samples are digested in vitro with a standardized rumen inoculum procedure. Starch digestion is
determined by difference in starch content between intact and digested samples.		Heuer, C.H., J.P. Goeser, and R.D. Shaver. 2013. Starch digestion variation between in vitro and in situ
digestion techniques. J Dairy Sci. E Suppl. (2013 JAM) abstract.
Richards, C.J., J.F. Pedersen, R.A. Britton, R.A. Stock and C.R. Krehbiel. 1995. In vitro starch
disappearance procedure modifications. Anim Feed Sci Tech. 55: 35-45.
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Kernel Processing ScoreChopped corn plant samples are subsampled, dried, and sieved aggressively to separate large (>4.75mm), medium (1.18 to 4.75mm), and fine (<1.18mm) particles. The three fractions of the sample are ground and analyzed separately to determine starch content. The starch percent that passes through 4.75mm screen is determined and used to quantify kernel processing score.Ferreira, G. 2002. Nutrient evaluation corn silage: Chemical and Physical characteristics of corn silage and their effects on in vitro disappearance. M.S. Thesis (Ch. 3) Univ. of Wisconsin-Madison.

Ferreira, G., and D.R. Mertens. 2005. Chemical and physical characteristics of corn silages and their effects of in vitro disappearance. J. Dairy Sci 88:4414-4425.

Mertens, D.R. 2005. Particle size, fragmentation index, and effective fiber: Tools for evaluating the physical attributes of corn silages. Proc. 2005 Four-State Dairy Nutrition. And Management Conf. pg. 211-220.
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LigninAnkom Technologies Method 8https://www.ankom.com/sites/default/files/document-files/Method_8_Lignin_in_beakers.pdf
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MineralsSamples are digested at 120 ०C in acid to extract minerals. Dilute acid solutions are analyzed by ICP-OES to determine the content of Ca, Mg, K, P, Zn, B, Cu, S, Na, Mn, and Fe simultaneously.Modified AOAC Official Method 968.08 - Minerals in Animal Feed and Pet Food
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Mold and Yeast Samples are plated at 1:1000, 1:10,000, and 1:100,000 dilution rates on 3 potato dextrose agar plates using Butterfield Buffer as diluent. Plates are incubated at 25०C for 5 days and read on the lowest dilution plate with <100 colony forming units(CFU). Results are reported as CFU/g. BAM R11: Butterfield’s Phosphate-buffered dilution water. 2013. FDA Laboratory Methods. Accessed online at http://www.fda.gov/Food/FoodScienceResearch/LaboratoryMethods/ucm061208.htm.
Compendium of methods for the microbiological examination of foods, 4th Ed. 2001. American Public Health Assoc., Washington, D.C.
Handbook of Food Spoilage Yeasts, 2nd Ed. 2007. Deak ed. CRC Press, Boca Raton, FL.
Introduction to Food-Born Fungi. 3rd Ed. 1988. Samson, R.A., and E.S. Reenen-Hoekstra. Centraalburea voor Schimmelcultures, BAARN. The Netherlands.
Adesogan, A.T., N. Krueger, M.B. Salawu, D.B. Dead and C.R. Staples. 2004. The influence of treatment with dual purpose bacterial inoculants or soluble carbohydrates on the fermentation and aerobic stability of bermudagrass. J Dairy Sci. 87:3407-3416.

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MSPE (Multi-Step Protein Evaluation)In vitro protein digestion assay developed at Cornell University https://hdl.handle.net/1813/36488
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Neutral Detergent FiberAnkom Technologies Method 6https://www.ankom.com/sites/default/files/document-files/Method_6_NDF_A200.pdf
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NIRAfter grinding, the sample is equilibrated to room temperature, and a subsample is packed into a Foss Ring Cup or equivalent for the platform in use. Samples are scanned on a Foss System II model 5000, Foss DS2500, or BlueSun Phoenix 5000. Spectral data between 1100 and 2500 nm is recorded and used for prediction.
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NitrateSample is extracted using 2M KCL solution, shaken for 15 minutes, and filtered. Extract is analyzed  by Skalar SAN ++ Segmented Flow SA 5000 analyzer. 

Skalar Method: Automated procedure  based on the modified cadmium reduction reaction method; After dialysis, the sample is buffered at pH 8.2 and is passed through a column containing granulated copper-cadmium to reduce the nitrate to nitrite.  The total nitrite is determined by diazotizing with sulfanilamide and coupling with N-(1-naphthyl)ethylenediamine dihydrochloride to form a highly colorized azo dye which is read at 540 nm		Skalar Method Resources: Standard method for examination of water and wastewater, 15th edition, 1980.
Navone, R., 1964, Proposed method for nitrate in potable waters, Amer. J., Water works Ass. 56:781, 1964.Walinga, I., van Vark, W., Houba, V.J.G., van der Lee, J.J., Plant Analysis Procedure, Part 7, Department of Soil Science and Plant Nutrition, Wageningen Agricultural University, Syllabus 1989, page 197-200.
ISO 13395:1996, Determination of nitrite nitrogen and nitrate nitrogen and the sum of both by flow analysis (CFA) and spectrometric detection. 
ISO 3696:1987, Water for analytical laboratory use. Specification and test methods. 
ASTM, D1193, Standard Specification for Reagent Water. 
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Soluble Crude ProteinA dried, ground sample is extracted with a borate-phosphate buffer pH 6.75 at 39 degrees C for 1 hour. After the hour the sample is filtered and soluble crude protein in the supernatant is determined using CP procedure.G. Licitra, T.M. Hernandez, P.J. Van Soest, 1996. Standardization of procedures for nitrogen fractionation of ruminant feeds. Animal Feed Science and Technology,Volume 57, Issue 4.
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Starch(Hall, MB. 2009)Hall, MB. 2009. Determination of Starch, Including Maltooligosaccharides, in Animal Feeds: Comparison of Methods and a Method Recommended for AOAC Collaborative Study. Journal of AOAC International. 92:42-49.
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Toxins (in House)HPLC MS/MS
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uNDF/NDFD1mm ground samples are digested in triplicate in vitro with a standardized rumen inoculum at 39 °C under continuous carbon dioxide. In vitro samples are frozen or refluxed immediately to terminate fermentation. The NDF content of completed samples is determined using a neutral detergent solution containing alpha amylase and sodium sulfite as described by Goering and Van Soest. Samples are vacuum filtered using Ahlstrom 141 7 cm Glass Microfiber filters.

Goeser, JP. 2008. Improvement of rumen in vitro NDF digestion techniques and data analysis. PhD Thesis. University of Wisconsin – Madison.

Goeser, JP and DK Combs. 2009. An alternative method to assess 24-h ruminal in vitro NDF digestibility. J Dairy Sci. 92:3833-3841.

Goeser, JP, PC Hoffman, and DK Combs. 2009. Modification of a rumen fluid priming technique for measuring in vitro NDF digestibility. J Dairy Sci. 92:3842-3848.

Goeser, J., C. Heuer, and L. Meyer. 2013. In vitro NDF digestion parameters differ when using a Forage Fiber Bag. J Dairy Sci. E Suppl. (2013 JAM) abstract

Goering, H. K., and P. J. Van Soest. 1970. Forage Fiber Analyses (Apparatus, Reagents, Procedures, and Some Applications). Agric. Handbook No. 379. ARS-USDA, Washington, DC.
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TMRD Assay (uNDF 120)1mm ground samples are weighed into acetone washed Ankom 57 fiber bags, sealed, and digested in triplicate in vitro with a standardized rumen inoculum at 39 °C under continuous carbon dioxide. After 120h, bags are removed and rinsed under cold water until the water runs clear. Bags are either run immediately for NDF or dried at 55 °C and then run for NDF. The NDF content of completed samples is determined using a neutral detergent solution containing alpha amylase and sodium sulfite as described by Ankom Technologies Method 6. uNDF 120 is used as an internal marker to calculate apparent nutrient digestibility as described in Schalla et al., 2012. Schalla, A., Meyer, L., Meyer, Z., Onetti, S., Schultz, A., & Goeser, J. (2012). Hot topic: Apparent total-tract nutrient digestibilities measured commercially using 120-hour in vitro indigestible neutral detergent fiber as a marker are related to commercial dairy cattle performance. Journal of Dairy Science, 95(9), 5109–5114. https://doi.org/10.3168/jds.2012-5650

https://www.ankom.com/sites/default/files/document-files/Method_6_NDF_A200.pdf
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VFAVolatile fatty acids are extracted from the sample in a 1:10 ratio of sample and deionized water. The extract is centrifuged and the supernatant is combined with calcium hydroxide and copper sulfate, centrifuged again, and the supernatant is analyzed by HPLC equipped with a reverse-phase ion exclusion column and a refractive index detector.Method was developed in conjunction with Dr. Richard Muck of USDA-ARS, Madison, WI
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WSC(Dubois et al., 1956; Hall, 2013)Dubois, M., K.A. Gilles, J.K. Hamilton, P.A. Rebers, and F. Smith. 1956. Colorimetric method for
determination of sugars and related substances. Anal. Chem. 28:350-356.
Hall, M.B. 2013. Efficacy of reducing sugar and phenol-sulfuric acid assays for analysis of soluble
carbohydrates in feedstuffs. Animal Feed Science and Technology. 185:94-100.
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Crude Fat (acid hydrolysis)Ankom Technology (Total Fat by Acid Hydrolysis)https://www.ankom.com/sites/default/files/document-files/HCl_Analytical_Proc.pdf
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Crude Fat (ether extract)Ankom Technology (Modified AOCS Am5-04)https://www.ankom.com/sites/default/files/document-files/Crude_Fat_Abstract.pdf
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pHSample is mixed with deionized water and read with a combination pH electrode
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Notes/ Updates:
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Chloride 2/6/262/6/26 addition would like an AOAC method
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